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. 2014 Feb 10;23(12):1392–1404. doi: 10.1089/scd.2013.0528

FIG. 5.

FIG. 5.

TGFβ1 released from the diseased aorta induces the migration of MSCs to the aortic lesions. (A) Immunohistochemical staining of active TGFβ1 in aorta of ldlr−/− mice fed CHD or HFD for 1 or 2 weeks. (B) Numbers of migrated MSCs in Aorta CM-based migration assays using CHD CM or HFD CM with 10 μg/mL neutralizing TGFβ antibody (1D11) or same concentration of control antibody (13C4) were counted per field of view (FV,×20 magnification). n=4. *P<0.01. (C) Numbers of migrated cells in Aorta CM-based migration assays using HFD CM with addition of individual inhibitors (2 μM SB505124, 10 μM Y27632, 10 μM SB202190, 10 μM U0126, and 10 μM SP600125). Numbers of migrated cells and differentiated cells were counted per field of view. n=4. *P<0.001, versus vehicle control. (D) MSCs were incubated with CHD CM or HFD CM with addition of 10 μg/mL control IgG antibody (13C4) or TGFβ neutralizing antibody (1D11) together with vehicle or 2 μM SB505124 as indicated for 2 h. Protein was extracted from MSCs, and western blot analysis was conducted using antibodies against phosphorylated Smad2/3 (p-Smad2/3) or total Smad2/3, respectively. (E) Alizarin Red staining of the MSCs incubated with control DMEM, CHD CM, or HFD CM with addition of 10 μg/mL neutralizing TGFβ antibody (1D11) or same concentration of control antibody (13C4). TGFβ1, transforming growth factor.